LOCUS       sequence-14202-e        4774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4774)
  AUTHORS   Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller 
            JA, Somia NV, Bogdanove AJ, Voytas DF
  TITLE     Efficient design and assembly of custom TALEN and other TAL 
            effector-based constructs for DNA targeting.
  JOURNAL   Nucleic Acids Res. 2011 Jul;39(12):e82. doi: 10.1093/nar/gkr218
  PUBMED    21493687
REFERENCE   2  (bases 1 to 4774)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2011-07"; volume: "39(12):e82. doi"; pages: " 
            10.1093/nar/gkr21"
COMMENT     Sequence Label: pTAL2
FEATURES             Location/Qualifiers
     source          1..4774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(927..1334)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     1199..1215
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1225..1243
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1291..1311)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1291..1309)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1330..1346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1343..1365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1354..1370
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1378..1384,1385..1402,1403..1408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1423..1444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(2282..2381)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2399..2417)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2422..2438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(2600..3391,3392..3460))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3223..3242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3588..4176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4077..4096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4330..4347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4506..4533)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4625..4711)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccgaattc gcccttttgc atgtaaatag gaggtgcacc atggatccca
      181 ttcgtccgcg caggccaagt cctgcccgcg agcttctgcc cggaccccaa ccggataggg
      241 ttcagccgac tgcagatcgt ggggtgtctg cgcctgctgg cagccctctg gatggcttgc
      301 ccgctcggcg gacggtgtcc cggacccggc tgccatctcc ccctgcgccc tcacctgcgt
      361 tctcggcggg cagcttcagc gatctgctcc gtccgttcga tccgtcgctt cttgatacat
      421 cgcttcttga ttcgatgcct gccgtcggca cgccgcatac agcggctgcc ccagcagagt
      481 gggatgaggc gcaatcggct ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg
      541 ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac
      601 cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac gctcggctac agtcagcagc
      661 agcaagagaa gatcaaaccg aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg
      721 tgggccatgg gtttacacac gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag
      781 ggaccgtcgc tgtcacgtat cagcacataa tcacggcgtt gccagaggcg acacacgaag
      841 acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg
      901 atgcggggga gttgagaggt ccgccgttac agttggacac aggccaactt gtgaagattg
      961 caaaacgtgg cggcgtgacc gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg
     1021 gtgcccccct ggagacgggc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     1081 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     1141 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     1201 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg
     1261 ccccccctcg aggtcctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     1321 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1441 attaattgcg ttgcgctcac tgcccgcttt ccaccggtcg tctccaacga ccacctcgtc
     1501 gccttggcct gcctcggcgg acgtcctgcc atggatgcag tgaaaaaggg attgccgcac
     1561 gcgccggaat tgatcagaag agtcaatcgc cgtattggcg aacgcacgtc ccatcgcgtt
     1621 gccgactacg cgcaagtggt tcgcgtgctg gagtttttcc agtgccactc ccacccagcg
     1681 tacgcatttg atgaggccat gacgcagttc gggatgagca ggaacgggtt ggtacagctc
     1741 tttcgcagag tgggcgtcac cgaactcgaa gcccgcggtg gaacgctccc cccagcctcg
     1801 cagcgttggg accgtatcct ccaggcatca gggatgaaaa gggccaaacc gtcccctact
     1861 tcagctcaaa caccggatca ggcgtctttg catgcattcg ccgattcgct ggagcgtgac
     1921 cttgatgcgc ccagcccaat gcacgaggga gatcagacgc gggcaagcag ccgtaaacgg
     1981 tcccgatcgg atcgtgctgt caccggcccc tccgcacagc aggctgtcga ggtgcgcgtt
     2041 cccgaacagc gcgatgcgct gcatttgccc ctcagctgga gggtaaaacg cccgcgtacc
     2101 aggatctggg gcggcctccc ggatcctggt acgcccatgg ctgccgacct ggcagcgtcc
     2161 agcaccgtga tgtgggaaca agatgcggac cccttcgcag gggcagcgga tgatttcccg
     2221 gcattcaacg aagaggaact cgcatggttg atggagctat tgcctcagaa gggcgaattc
     2281 gacccagctt tcttgtacaa agttggcatt ataaaaaata attgctcatc aatttgttgc
     2341 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     2401 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     2461 aatctctgat gttacattgc acaagataaa aatatatcat catgagatta tcaaaaagga
     2521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3481 attgaagcat ttatcagggt tattgtctca tgaccaaaat cccttaacgt gagttacgcg
     3541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3721 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3961 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4201 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4261 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4321 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4381 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4441 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     4501 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     4561 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     4621 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     4681 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     4741 acgctagcat ggatgttttc ccagtcacga cgtt
//
